RESUMEN
Inhibition of the 26S proteasome is an attractive approach for anticancer therapy. Proteasome inhibitors are known to selectively target cancer cells and make them more sensitive to chemotherapeutic agents. Murraya koenigii is a medicinally important herb of Asian origin and a rich source of bioactive compounds such as flavonoids and alkaloids. In the present study, we investigated the proteasome inhibitory and apoptotic effect of M. koenigii leaf extract in vivo in a xenograft tumor mouse model, and also assessed the toxicity if any in normal mice. M. koenigii extract did not lead to any toxicity in mice. Analysis of extract revealed the presence of flavonoid compounds which act as proteasome inhibitors. Quercetin treatment led to the decrease in the cell viability and arrest of cells in G2/M phase. Quercetin, Apigenin, Kaempferol and Rutin; flavonoids present in the leaf extract, dose-dependently inhibited the endogenous 26S proteasome activity in MDA-MB-231 cells. Reduction in tumor growth was associated with a decrease in proteasomal enzyme activities in the treated groups. Increased caspase-3 activity and TUNEL-positive cells indicated enhanced apoptosis with Murraya leaf extract treatment. Decreased expression of angiogenic and anti-apoptotic gene markers is indicative of inhibition of angiogenesis and promotion of apoptosis in the leaf extract treated tumors.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Flavonoides/farmacología , Murraya/química , Extractos Vegetales/farmacología , Complejo de la Endopetidasa Proteasomal/metabolismo , Inhibidores de Proteasoma/farmacología , Animales , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/aislamiento & purificación , Neoplasias de la Mama/patología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Flavonoides/química , Flavonoides/aislamiento & purificación , Humanos , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Neoplasias Mamarias Experimentales/patología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Hojas de la Planta/química , Inhibidores de Proteasoma/química , Inhibidores de Proteasoma/aislamiento & purificación , Relación Estructura-ActividadRESUMEN
Skin cancer is among the most common cancers worldwide and identifiable molecular changes for early and late stage of skin tumorigenesis can suggest the better targets for its control. In this study, we investigated the status of K-Ras-PI3K-AKTpathway followed by NF-κB, cyclin D1, MMP-9 and regulatory micro RNA during 7, 12-dimethylbenz[a]anthracene (DMBA) induced mouse skin tumorigenesis and its prevention by butyric acid (BA), nicotinamide (NA) and calcium glucarate (CAG), individually or in combination with respect to time. DMBA upregulated the K-Ras, PI3K, Akt, NF-κB, cyclin D1 and MMP-9, but downregulated the PTEN in a time dependent manner. DMBA also reduced the levels of micoRNA let-7a but induced the levels of miR-21 and miR-20a as a function of time. BA, NA and CAG were found to prevent DMBA induced changes, but they were most effective when used together in a combination. Reduced let-7a and miR-211 were correlated with the overexpression of K-Ras and MMP-9. Overexpression of miR-21 and miR-20a was correlated with the down regulation of PTEN and overexpression of Cyclin D1. Collectively, the enhanced chemopreventive potential of natural compound in combination via regulation of K-Ras-PI3K-AKTpathway along with regulatory micro RNAs provide a newer and effective mean for cancer management.
Asunto(s)
Ácido Butírico/farmacología , Ácido Glucárico/farmacología , MicroARNs/genética , Niacinamida/farmacología , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Neoplasias Cutáneas/metabolismo , 9,10-Dimetil-1,2-benzantraceno , Animales , Antineoplásicos/farmacología , Western Blotting , Ratones , Fosfatidilinositol 3-Quinasas/genética , Proteínas Proto-Oncogénicas c-akt/genética , Neoplasias Cutáneas/inducido químicamenteRESUMEN
We explored the basis of the combinatorial chemopreventive effect of butyric acid (BA), nicotinamide (NA) and calcium glucarate (CAG) on mouse skin exposed to 7,12-dimethylbenz(a)anthracene (DMBA). We studied the effects of topical application of DMBA in the presence or absence of BA, NA and CAG on the regulators of apoptosis. DMBA treatment suppressed Bax, Bax/Bcl-2 ratio, release of cyt c, Apaf1, caspase-9, -3 mediated apoptosis. Downregulation of p21 and upregulation of Bcl-2, mut p53 were also observed in only DMBA treated mice. Simultaneous application of BA, NA and CAG induced a mitochondria-mediated apoptosis, characterized by a rise in the Bax, Bax/Bcl-2 ratio, release of cyt c, upregulation of Apaf1 with down-stream activation of caspase-9, -3. Furthermore treatment with BA, NA and CAG demonstrated an upregulation of p21 and downregulation of Bcl-2, mut p53. But this effect was enhanced in the presence of all the three compounds together in combination. Chemoprevention by a combination of BA, NA and CAG by inducing the apoptosis, the natural cell death, suggest the importance of the potential combinational strategies capable of preventing skin tumor development.
Asunto(s)
9,10-Dimetil-1,2-benzantraceno/toxicidad , Anticarcinógenos/farmacología , Apoptosis/efectos de los fármacos , Productos Biológicos/farmacología , Carcinogénesis/efectos de los fármacos , Neoplasias Cutáneas/inducido químicamente , Neoplasias Cutáneas/prevención & control , Animales , Factor Apoptótico 1 Activador de Proteasas/genética , Factor Apoptótico 1 Activador de Proteasas/metabolismo , Ácido Butírico/farmacología , Caspasas/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Citocromos c1/metabolismo , Citoplasma/efectos de los fármacos , Citoplasma/metabolismo , Interacciones Farmacológicas , Activación Enzimática/efectos de los fármacos , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Ácido Glucárico/farmacología , Ratones , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Niacinamida/farmacología , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Neoplasias Cutáneas/metabolismo , Neoplasias Cutáneas/patología , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismoRESUMEN
The vitamin D endocrine system is functional in the adipose tissue, as demonstrated in vitro, in cultured adipocytes, and in vivo in mutant mice that developed altered lipid metabolism and fat storage in the absence of either 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] or the vitamin D receptor. The aim of the present study was to examine the role of vitamin D and calcium on body adiposity in a diet-induced vitamin D deficient rat model. Vitamin D-deficient rats gained less weight and had lower amounts of visceral fat. Consistent with reduced adipose tissue mass, the vitamin D-deficient rats had low circulating levels of leptin, which reflects body fat stores. Expression of vitamin D and calcium sensing receptors, and that of genes involved in adipogenesis such as peroxisome proliferator-activated receptor, fatty acid synthase and leptin were significantly reduced in white adipose tissue of deficient rats compared to vitamin D-sufficient rats. Furthermore, the expression of uncoupling proteins (Ucp1 and Ucp2) was elevated in the white adipose tissue of the deficient rat indicative of higher energy expenditure, thereby leading to a lean phenotype. Expression of the p160 steroid receptor coactivator3 (SRC3), a key regulator of adipogenesis in white adipose tissue was decreased in vitamin D-deficient state. Interestingly, most of the changes observed in vitamin D deficient rats were corrected by calcium supplementation alone. Our data demonstrates that dietary vitamin D and calcium regulate adipose tissue function and metabolism.
Asunto(s)
Adiposidad/fisiología , Canales Iónicos/genética , Proteínas Mitocondriales/genética , Coactivador 3 de Receptor Nuclear/genética , Deficiencia de Vitamina D/metabolismo , Adipogénesis/genética , Adiponectina/sangre , Tejido Adiposo/metabolismo , Animales , Calcio/farmacología , Dieta , Acido Graso Sintasa Tipo I/genética , Expresión Génica , Hipocalcemia/etiología , Hipocalcemia/genética , Hipocalcemia/metabolismo , Insulina/sangre , Leptina/sangre , Leptina/genética , Lípidos/sangre , Hígado/metabolismo , Masculino , Coactivador 3 de Receptor Nuclear/metabolismo , PPAR gamma/genética , Ratas Sprague-Dawley , Proteína Desacopladora 1 , Proteína Desacopladora 2 , Deficiencia de Vitamina D/complicaciones , Deficiencia de Vitamina D/genéticaRESUMEN
BACKGROUND: The study was undertaken to provide value addition to spent eri silkworm as an alternative source of edible oil for the food and feed industry by carrying out a short-term nutritional and toxicological evaluation of eri silkworm pupae oil using Wistar NIN rats. RESULTS: Growth performance of rats fed either sunflower oil (Control) or eri silkworm pupae oil (Experimental) was comparable. Histopathological examination of the various tissues showed no signs of toxicity even after feeding the eri silkworm oil for 18 weeks. Serum cholesterol and triglyceride was significantly reduced (P < 0.05) while high-density lipoprotein cholesterol was significantly increased (P < 0.05) which is attributed to the high α-linolenic acid content of eri silkworm oil. CONCLUSION: The study showed that eri silkworm pupae oil is safe and nutritionally equivalent to commonly used vegetable oils. Eri silkworm pupae can be harvested to provide a cost effective alternative edible oil that can be used to nutritional advantage in the food and feed industry. Therefore eri silkworm and its host plants offer an excellent example of multiple product crops and of sustainable agricultural practice with excellent opportunity for economic and nutritional benefits.
Asunto(s)
Bombyx/química , Colesterol/sangre , Grasas de la Dieta/farmacología , Crecimiento/efectos de los fármacos , Aceites/farmacología , Triglicéridos/sangre , Ácido alfa-Linolénico/farmacología , Animales , HDL-Colesterol/sangre , Grasas de la Dieta/análisis , Femenino , Masculino , Valor Nutritivo , Aceites/efectos adversos , Aceites/química , Pupa/química , Ratas , Ratas Wistar , Ácido alfa-Linolénico/análisisRESUMEN
The enzyme 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) amplifies intracellular glucocorticoid action by converting inactive glucocorticoids to their active forms in vivo. Adipose-specific overexpression of 11ß-HSD1 induces metabolic syndrome in mice, whereas 11ß-HSD1 null mice are resistant to it. Dietary trans and saturated fatty acids (TFAs and SFAs) are involved in the development of metabolic syndrome, whereas polyunsaturated fatty acids (PUFA) offer protection against this. Here, we report the effects of chronic feeding of different diets containing vanaspati (TFA rich), palm oil (SFA rich) and sunflower oil (PUFA rich) at 10%level on 11ß-HSD1 gene expression in rat retroperitoneal adipose tissue. 11ß-HSD1 gene expression was significantly higher in TFA rich diet-fed rats compared to SFA rich diet-fed rats, which in turn was significantly higher than PUFA rich diet-fed rats. Similar trend was observed in the expression of CCAAT-enhancer binding protein-α (C/EBP-α), the main transcription factor required for the expression of 11ß-HSD1. We propose that TFAs and SFAs increase local amplification of glucocorticoid action in adipose tissue by upregulating 11ß-HSD1 by altering C/EBP-α-gene expression. The increased levels of glucocorticoids in adipose tissue may lead to development of obesity and insulin resistance, thereby increasing the risk of developing metabolic syndrome.